LOCUS       40924_28087        7413 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLeo645, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7413)
  AUTHORS   Scheller L, Strittmatter T, Fuchs D, Bojar D, Fussenegger M.
  TITLE     Generalized extracellular molecule sensor platform for programming 
            cellular behavior
  JOURNAL   Nat. Chem. Biol. 14 (7), 723-729 (2018)
  PUBMED    29686358
REFERENCE   2  (bases 1 to 7413)
  AUTHORS   Scheller L, Strittmatter T.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-NOV-2017) D-BSSE, ETH Zurich, Mattenstrasse 26, Basel 
            4058, Switzerland
REFERENCE   3  (bases 1 to 7413)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7413)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. Chem. 
            Biol."; date: "2018"; volume: "14"; issue: "7"; pages: "723-729"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-NOV-2017) D-BSSE, ETH Zurich, Mattenstrasse 26, Basel 4058, 
            Switzerland"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7413
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    7..215
                     /label=Psv40
                     /note="Psv40"
     promoter        18..214
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      65..200
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      240..249
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    246..308
                     /label=igk secretion signal
                     /note="igk secretion signal"
     misc_feature    246..294
                     /label=secrete
                     /note="secrete"
     misc_feature    309
                     /label=scfv
                     /note="scfv"
     misc_feature    315..692
                     /label=anti RR120
                     /note="anti RR120"
     misc_feature    705..1448
                     /label=mouse EpoR-D1_D2_TM
                     /note="mouse EpoR-D1_D2_TM"
     misc_feature    978..980
                     /label=F93A
                     /note="F93A"
     misc_feature    1380..1448
                     /label=transmembrane
                     /note="transmembrane"
     misc_feature    1461..2732
                     /label=mFGFR 405-end
                     /note="mFGFR 405-end"
     polyA_signal    2786..3010
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3056..3484
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3498..3827
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3894..4685
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    4862..4995
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5032..5048)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5056..5072)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5080..5110)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5125..5146)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5434..6019)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6193..7050)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7051..7155)
                     /label=AmpR promoter
ORIGIN
        1 acgcgtgatc tgcgatctgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
       61 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatc gctgactaat
      121 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
      181 aggaggcttt tttggaggcc taggcttttg caaaaagctt cgaatcgcgc tagcggccgg
      241 ccaccatgga aactgatact ttgctgctct gggtcctgct gctgtgggtc cctggatcaa
      301 cgggggacgg atccgtgcag ttgcaagaat cgggtggtgg actggttcaa gccggtgata
      361 gtttgaaact gtcatgcgaa gccagtggcg attccattgg aacttatgtc attggttggt
      421 ttcgccaagc accggggaaa gagcgcattt accttgccac tattggtcgt aacctggtcg
      481 ggccgtccga tttttataca cgctacgctg actctgtaaa gggtcgcttc gcagtgagtc
      541 gtgacaacgc caagaacact gtaaacttac agatgaactc tcttaaacct gaagatactg
      601 ctgtttacta ttgtgctgcg aaaacgacaa cgtggggggg aaacgatcct aataactgga
      661 attattgggg acaaggcact caggtgaccg tatccggaga attcgcacct tcacccagcc
      721 tcccggaccc caagtttgag agcaaagcgg ccctgctggc atcccggggc tccgaagaac
      781 ttctgtgctt cacccaacgc ttggaagact tggtgtgttt ctgggaggaa gcggcgagct
      841 ccgggatgga cttcaactac agcttctcat accagctcga gggtgagtca cgaaagtcat
      901 gtagcctgca ccaggctccc accgtccgcg gctccgtgcg tttctggtgt tcactgccaa
      961 cagcggacac atcgagtgct gtgccgctgg agctgcaggt gacggaggcg tccggttctc
     1021 ctcgctatca ccgcatcatc catatcaatg aagtagtgct cctggacgcc cccgcggggc
     1081 tgctggcgcg ccgggcagaa gagggcagcc acgtggtgct gcgctggctg ccacctcctg
     1141 gagcacctat gaccacccac atccgatatg aagtggacgt gtcggcaggc aaccgggcag
     1201 gagggacaca aagggtggag gtcctggaag gccgcactga gtgtgttctg agcaacctgc
     1261 ggggcgggac gcgctacacc ttcgctgttc gagcgcgcat ggccgagccg agcttcagcg
     1321 gattctggag tgcctggtct gagcccgcgt cactactgac cgctagcgac ctggaccctc
     1381 tcatcttgac gctgtctctc attctggtcc tcatctcgtt gttgctgacg gttctggccc
     1441 tgctgtccgc cgctgcggcg atgaagagcg gcaccaagaa gagcgacttc catagccaga
     1501 tggctgtgca caagctggcc aagagcatcc ctctgcgcag acaggtaaca gtgtcagctg
     1561 actccagtgc atccatgaac tctggggttc tcctggttcg gccctcacgg ctctcctcca
     1621 gcgggacccc catgctggct ggagtctccg aatatgagct ccctgaggat ccccgctggg
     1681 agctgccacg agacagactg gtcttaggca aaccacttgg cgagggctgc ttcgggcagg
     1741 tggtgttggc tgaggccatc gggctggata aggacaaacc caaccgtgtg accaaagtgg
     1801 ccgtgaagat gttgaagtcc gacgcaacgg agaaggacct gtcggatctg atctcggaga
     1861 tggagatgat gaaaatgatt gggaagcaca agaatatcat caaccttctg ggagcgtgca
     1921 cacaggatgg tcctctttat gtcattgtgg agtacgcctc caaaggcaat ctccgggagt
     1981 atctacaggc ccggaggcct cctgggctgg agtactgcta taaccccagc cacaaccccg
     2041 aggaacagct gtcttccaaa gatctggtat cctgtgccta tcaggtggct cggggcatgg
     2101 agtatcttgc ctctaagaag tgtatacacc gagacctggc tgctaggaac gtcctggtga
     2161 ccgaggataa cgtaatgaag atcgcagact ttggcttagc tcgagacatt catcatatcg
     2221 actactacaa gaaaaccacc aacggccggc tgcctgtgaa gtggatggcc cctgaggcgt
     2281 tgtttgaccg gatctacaca caccagagcg atgtgtggtc ttttggagtg ctcttgtggg
     2341 agatcttcac tctgggtggc tccccatacc ccggtgtgcc tgtggaggaa cttttcaagc
     2401 tgctgaagga gggtcatcga atggacaagc ccagtaactg taccaatgag ctgtacatga
     2461 tgatgcggga ctgctggcat gcagtgccct ctcagagacc tacgttcaag cagttggtgg
     2521 aagacctgga ccgcattgtg gccttgacct ccaaccagga gtatctggac ctgtccatac
     2581 cgctggacca gtactcaccc agctttcccg acacacggag ctccacctgc tcctcagggg
     2641 aggactctgt cttctctcat gagccgttac ctgaggagcc ctgtctgcct cgacacccca
     2701 cccagcttgc caacagtgga ctcaaacggc gctaggcggc cgctcgagtc tagagggccc
     2761 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     2821 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     2881 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     2941 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     3001 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     3061 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3121 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3181 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3241 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3301 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3361 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3421 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3481 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3541 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3601 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3661 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3721 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     3781 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     3841 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     3901 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3961 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4021 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     4081 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4141 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4201 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4261 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4321 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4381 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     4441 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4501 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4561 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4621 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4681 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     4741 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4801 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4861 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4921 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4981 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     5041 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5101 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5161 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5221 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5281 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5341 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5401 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5461 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5521 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5581 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5641 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5701 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5761 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5821 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5881 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5941 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg
     6001 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6061 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6121 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6181 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6241 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6301 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6361 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6421 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6481 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6541 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6601 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6661 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6721 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6781 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6841 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6901 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6961 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7021 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7081 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7141 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     7201 tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     7261 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     7321 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
     7381 tttgcgctgc ttcgcgatgt acgggccaga tat
//
